Abstract
Purpose :
We have previously shown that Polycaprolactone (PCL)/Poly(glycerol sebacate) (PGS) nanofiber matrices show properties for ocular surface reconstruction and the possibility to introduce linker groups for drug attachment. To evaluate the biocompatibility of these materials, we now evaluated the biological activity of two human corneal cell types on these matrices.
Methods :
The nanofiber matrices were electrospun in different blends of PCL with PGS and with chitosan. Electrospun fiber meshes with either random orientation or aligned have been collected on a substrate and cut into circular samples with a diameter of 14 mm. After rinsing the samples in PBS buffer overnight and sterilization by UV-Light the samples were put into 24-well-Plates and incubated with HCE and HCK cells for 8 days. Wells with PCL foil were used as controls. During cultivation biological activities were measured for each sample after 2, after 5 and after 8 days by cell proliferation assay (WST-8). After the incubation time the samples were prepared for confocal microscopy and pictures have been taken to check the cell morphology. (Results shown as random: Mean ± SD / aligned: Mean ± SD) (AU=Absorbance Units)
Results :
HCE and HCK cells could be cultured on each of the materials, PCL alone, PCL blended with PGS and PCL blended with chitosan. On aligned fiber meshes an orientation of the HCE and the HCK cells in direction of the fibers can be detected on confocal microscopic pictures by checking the growth direction of the F-Actin fibers. In contrast the cells grew random on the random fiber meshes. The bioactivity of the HCE cells was lowest on PCL chitosane [0,006 ± 0,003 AU / 0,046 ± 0,02 AU] and similar on the fibers with PGS [0,175 ± 0,12 AU / 0,078 ± 0,03 AU] and PCL alone [0,149 ± 0,07 AU / 0,078 ± 0,01 AU], while the bioactivity for the HCK cells was low on each material.
Conclusions :
Blending PCL with PGS shows good results for culturing HCE cells, wether further research has to be conducted on the chitosane material. Combined with the good alignment of the cells to the oriented fiber meshes the blend of PCL with PGS shows the best attributes for the further research with the goal to produce active surfaces for the tissue engineering while further research on the blend of PCL with chitosane is needed.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.